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Characterization of psychrotrophic microorganisms producing beta-galactosidase activities.

机译:表征产生β-半乳糖苷酶活性的精神营养微生物。

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Investigations of psychrotrophic microorganisms have been limited even though the dominant environment of the Earth is cold and enzymes with high activities at low temperatures could have commercial uses. We have isolated and characterized three psychrotrophic strains with beta-galactosidase activities. The isolates, B7, D2, and D5, were gram-positive, catalase-positive, obligate aerobes. Cells observed with a scanning electron microscope appeared as rods during the early stages of growth but became coccoid during the stationary phase. An analysis of the amino acid composition of the cell walls demonstrated the presence of lysine as the predominant diamino acid in all three isolates. The cell cycle morphology and cell wall composition suggest that the three isolates are members of the genus Arthrobacter. The beta-galactosidase activities in whole cells were labile when incubated at 40 degrees C and had temperature optima about 20 degrees C below that of the enzyme encoded by the lacZ gene of Escherichia coli. Electrophoresis of extracts from the isolates in nondenaturing polyacrylamide gels detected at least two protein bands that hydrolyzed 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal), suggesting the presence of beta-galactosidase isozymes.
机译:即使地球的主要环境是寒冷的,对精神营养微生物的研究也受到限制,并且在低温下具有高活性的酶可能具有商业用途。我们已经分离并表征了具有β-半乳糖苷酶活性的三种精神营养菌株。分离株B7,D2和D5为革兰氏阳性,过氧化氢酶阳性,专性需氧菌。用扫描电子显微镜观察到的细胞在生长的早期阶段以棒状出现,而在静止期变成球状。对细胞壁氨基酸组成的分析表明,在所有三个分离物中,赖氨酸是主要的二氨基酸。细胞周期的形态和细胞壁的组成表明这三个分离株是节杆菌属的成员。当在40°C孵育时,全细胞中的β-半乳糖苷酶活性不稳定,并且其最佳温度比大肠杆菌lacZ基因编码的酶低约20°C。在非变性聚丙烯酰胺凝胶中分离株的提取物进行电泳检测,发现至少有两个蛋白带水解了5-溴-4-氯-3-吲哚基-β-D-半乳糖吡喃糖苷(X-Gal),表明存在β-半乳糖苷酶同工酶。

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